Stemness CD24 activation promotes hepatocellular carcinoma progression via an immune escape mechanism

Table 1 Analysis of clinical data of patients with chronic liver diseases

Group	n	Male/female	Age (years)	HBsAg (+/-)	AFP (ng/mL)			Child-Pugh
					≤ 20	21-399	≥ 400	A	B	C
NC	111	57/56	22-79	0/111	111	0	0	-	-	-
CH	72	55/17	23-75	72/0	55	17	0	-	-	-
LC	60	40/20	32-88	60/0	42	17	1	48	9	3
HCC	129	93/36	37-86	97/32	38	53	38	97	23	9

AFP: Alpha-fetoprotein; NC: Normal control; CH: Chronic hepatitis; HCC: Hepatocellular carcinoma; HBsAg (+/-): Hepatitis B virus surface antigen (positive/negative); LC: Liver cirrhosis.

Table 2 Uni-/multi-variable analysis with several predictors in hepatocellular carcinoma patients

Variable	Univariate analysis			Multivariable analysis
	HR	P > |z|	95%CI	HR	P > |z|	95%CI
Age (years)
≤ 60 vs > 60	1.482	0.502	0.512-3.661
Gender
Male vs female	0.931	0.828	0.411-2.183
Grade of differentiation
Low vs high	1.316	0.515	0.571-3.545
Tumor number
Single vs multiple	1.105	0.779	0.428-2.806
Liver cirrhosis
Absent vs present	2.805	0.018a	1.161-7.531	2.401	0.013a	1.038-5.187
Hepatitis B virus
Absent vs present	0.422	0.015a	0.156-0.805	0.361	0.008a	0.152-0.796
Periportal embolus
Absent vs present	5.589	0.126	0.791-41.033
AFP (μg/L)
< 400 vs ≥ 400	0.788	0.402	0.516-1.328
TNM staging
I-II vs III-IV	0.221	0.187	0.134-1.485
CD24 expression
Low vs high	12.586	< 0.001a	3.531-52.023	5.58	< 0.001a	2.421-14.928

^aP < 0.05.

AFP: Alpha-fetoprotein; HCC: Hepatocellular carcinoma; TNM: Tumor-node-metastasis; HR: Hazard ratio; CD: Clusters of differentiation; 95%CI: 95% confidence interval.

Table 3 Clinicopathological characteristics of serum clusters of differentiation 24 levels in hepatocellular carcinoma (mean ± SD)

Group	n	CD24 (μg/L)	t value	P value
Age (years)
≤ 60	66	99.7 ± 27.5	0.467	0.641
> 60	63	97.3 ± 30.8
Gender
Male	93	100.8 ± 30.5	1.443	0.502
Female	36	92.6 ± 24.4
AFP (ng/mL)
< 400	91	98.9 ± 29.1	0.763	0.447
≥ 400	38	94.6 ± 29.4
HBV infection
Absent	32	88.0 ± 18.3	2.773	0.006
Present	97	103.7 ± 30.2
Tumor size (cm)
≤ 5	73	88.5 ± 21.4	4.795	< 0.001
> 5	56	111.4 ± 32.7
Gross classification
Unifocal	85	88.8 ± 19.9	6.096	< 0.001
Multifocal	44	117.4 ± 34.9
Periportal embolus
Absent	119	96.8 ± 28.9	2.358	0.02
Present	10	119.2 ± 28.3
Lymph node metastasis
Without	59	91.4 ± 20.4	2.675	0.008
With	70	104.8 ± 33.6
Differentiation
Poor	35	118.9 ± 35.5	5.364	< 0.001
Moderate & well	94	90.9 ± 22.1
Extra-hepatic metastasis
Without	97	94.6 ± 25.6	2.743	0.007
With	32	110.5 ± 35.8
TNM stage
I-II	71	91.8 ± 27.8	2.713	0.008
III-IV	58	106.8 ± 35.0
Child-Pugh classification
A	97	83.2 ± 25.1	5.592	< 0.001
B & C	32	114.7 ± 34.3

AFP: Alpha-fetoprotein; HBV: Hepatitis B virus; CD: Clusters of differentiation; TNM: Tumor-node-metastasis.

Table 4 Serum clusters of differentiation 24 and hepatocellular carcinoma markers in hepatocarcinogenesis (mean ± SD)

Marker	NC (n = 12)	Model rats in hepatocarcinogenesis
		Early (n = 17)	Middel (n = 15)	Late (n = 10)
CD24 (μg/L)	3.68 ± 0.63	7.28 ± 3.17a	10.61 ± 4.52b	14.33 ± 7.14b
ALT (IU/L)	14.75 ± 3.02	62.19 ± 29.79b	55.55 ± 26.80b	46.08 ± 13.39b
AST (IU/L)	15.88 ± 3.29	44.46 ± 9.25c	61.99 ± 18.21c	47.91 ± 21.22c
CD44 (μg/L)	5.32 ± 1.66	12.36 ± 8.18b	21.69 ± 5.38b	28.6 ± 9.14b
AFP (μg/L)	0.81 ± 0.16	1.25 ± 0.32	1.58 ± 0.51a	1.96 ± 0.75b
Wnt3a (μg/L)	0.96 ± 0.47	2.25 ± 2.24	6.28 ± 2.12b	9.98 ± 5.62c
GPC-3 (μg/L)	0.02 ± 0.08	1.01 ± 0.27	1.62 ± 0.42a	5.83 ± 1.54b

^aP < 0.05 vs the normal control (NC) group.

^bP < 0.01 vs the NC group.

^cP < 0.001 vs the NC group.

AFP: Alpha-fetoprotein; ALT: Alanine aminotransferase; AST: Aspartate aminotransferase; GPC-3: Glypican-3; CD: Cluster of differentiation.